Objective To investigate intraoperative neural monitoring(IONM)for prevention of recurrent laryngeal nerve injury(RLN)during thyroid cancer operation.Methods 288 cases of thyroid cancer admitted from Jun.2011 to Jun.2013 in our hospital were studied and they were divided into the observation group(140 cases) and the control group(148 cases) according to whether IONM was used during surgery.The incidence of RLN injury was compared between the 2 groups.Results The injury rate of RLN was lower in the observation group(0.67％) than in the control group(3.57％)and the operation time is shorter than the latter(P ＜0.001).There was no statistical difference for the operative bleeding volume and the average hospitalization time between the 2 groups (P ＞ 0.05).Conclusion INOM can help to shorten the operation time and reduce the incidence of RLN injury.

Objective The purpose of this study was to determine how to preserve the remaining pancreatic body and tail in the pancreatectomy. Methods In seven cases of pancreatectomy, three of them were the rupture of pancreatojejunal anastomosis, and four of them were the pancreatectomy for tumor in the pancreatic neck or body. During operations, a bridge internal drainages was used to drain the pancreatic juice into the adjacent jejunum. After the operations, the supportive treatment, continuous irrigation of peritoneal cavity and pancreatic enzyme inhibition were used. Results In all seven patients, the remaining pancreatic body and tail were successfully preserved. The endocrine functions of these patients recovered to nearly normal level and patients were discharged. Conclusions In preserving the remaining pancreatic body or tail, the bridge internal drainage has its advantage of convenience. It effectively preserves the exocrine of pancreas as well as its endocrine

Objective:To establish a presenilin enhancer-2 (PSENEN) gene-silenced human immortalized keratinocyte (HaCaT) cell model, and to evaluate the effect of PSENEN gene silencing on the proliferation of and γ-secretase expression in HaCaT cells.Methods:Three shRNAs targeting the PSENEN gene were constructed, and inserted into the linearized LV3-pGLV-h1-GFP-puro vector to establish a recombinant lentiviral expression plasmid. After restriction enzyme digestion and sequencing, lentiviral packaging and purification were performed, and lentiviral titer was determined. Cultured HaCaT cells were divided into 5 groups: shRNA1, shRNA2 and shRNA3 groups treated with the lentivirus solutions containing PSENEN gene-targeted shRNA1, shRNA2 and shRNA3 respectively, NC group treated with the lentivirus solution containing a negative control shRNA (shNC) , and blank group treated without lentivirus solution. After transfection, inverted fluorescence microscopy was performed, and transfection efficiency was determined by flow cytometry. Cell counting kit-8 (CCK8) assay was performed to evaluate the effect of PSENEN gene silencing on the proliferation of HaCaT cells, and real-time fluorescence-based quantitative PCR (qPCR) and Western blot analysis were conducted to determine the mRNA and protein expression of PSENEN, nicastrin (NCT) , presenilin-1 (PS1) and anterior pharynx defective 1a (APH1a) genes respectively. Statistical analysis was carried out by using repeated measures analysis of variance, one-way analysis of variance, and least significant difference t test for multiple comparisons. Results:Inverted fluorescence microscopy showed that fluorescence was observed in the shRNA1 group, shRNA2 group, shRNA3 group and NC group, and flow cytometry showed that the transfection efficiency was over 98% in the above 4 groups. qPCR and Western blot analysis revealed that the mRNA and protein expression of PSENEN gene significantly decreased in the shRNA1 (0.187 ± 0.010, 0.219 ± 0.097, respectively) , shRNA2 (0.163 ± 0.022, 0.208 ± 0.014, respectively) and shRNA3 (0.174 ± 0.009, 0.185 ± 0.062, respectively) groups compared with the NC group (1.054 ± 0.272, 1.076 ± 0.075, respectively, all P < 0.001) . CCK8 assay showed that the cellular proliferative activity significantly increased in the shRNA1 group compared with the NC group at 0, 12, 36 and 48 hours (all P < 0.05) , and there was no significant difference between the 2 groups at 24 or 60 hours (both P > 0.05) ; the cellular proliferative activity was significantly higher in the shRNA2 and shRNA3 groups than in the NC group at 0, 12, 24, 36, 48 and 60 hours (all P < 0.05) . There was no significant difference in the mRNA expression of NCT, PS1 and APH1a genes among the shRNA1 group, shRNA2 group, shRNA3 group, NC group, and blank group ( F= 8.168, 4.644, 1.981, respectively, all P > 0.05) , while the relative protein expression level of mature NCT (mNCT) , immature NCT (imNCT) , carboxyl-terminal fragment of PS1 (PS1-CTF) and APH1a significantly differed among the above 5 groups ( F= 39.268, 5.929, 27.842, 20.663, respectively, all P ≤ 0.01) . Compared with the NC group, the shRNA1, shRNA2 and shRNA3 groups all showed significantly decreased protein expression of mNCT, PS1-CTF and APH1a (all P < 0.01) , but insignificant changes in imNCT protein expression (all P > 0.05) . Conclusion:The PSENEN gene-silenced HaCaT cell model was successfully constructed, and the PSENEN gene silencing could lead to an increase in the cellular proliferative activity of HaCaT cells and a decrease in the protein expression of γ-secretase subunits mNCT, PS1-CTF and APH1a.

Objective To investigate the expression and relationship of livin and mutant p53 in hepatocellular carcinoma (HCC). Methods The expression of livin and mutant p53 were evaluated using the SP immunohistochemistry in 80 HCC tissue, 39 hepatic cirrhosis tissue, 33 normal tissue beside the hemangiomas of liver. Results The integrated A average results showed that the intension of positive expression of livin aligned by turns was HCC tissue (Median=5.09; P25～P75=3.06～8.28), hepatic cirrhosistissue(Median=3.05; P25～P75=2.49～4.25), normal liver tissue(Median=1.99; P25～P75=1.54～2.54) (P<0.001), respectively. It also showed that the intension of positive expression of p53 in HCC tissue (Median=43.13; P25～P75=20.41～78.53) was higher than that in hepatic cirrhosis tissue (Median=20.30;P25～P75=14.90～28.08), as well as in that of normal liver tissue (Median=15.52;P25～P75=12.81～21.80) (P <0.001), but it made no sense in statistics between hepatic cirrhosis tissue and normal tissue of liver. The expression of livin was obviously correlated with p53 in HCC tissue(r=0.241, P<0.05). Conclusion The overexpressian of livin and p53 and their positive correlation showed that livin may play a crucial role in the origin and development of HCC in coordination with p53.

Objective To study the correlation between lactate/albumin ratio level and incidence of multiple organ dysfunction syndrome (MODS) as well as mortality in patients with severe sepsis or septic shock.Methods In January 2012-September 2013, 54 patients who were admitted to the intensive care unit(ICU) of a hospital developed severe sepsis and septic shock on the first day of admission, clinical data of patients were analyzed.Results On the first and second days of admission, 30(55.56%)and 26(53.06%)patients developed MODS;lactate/albumin ratio between MODS group and non-MODS group on the first and second days of admission were both significantly different (both P<0.05).Multivariate logistic regression analysis showed that lactate/albumin ratio, oxygenation index (PaO2/FiO2), as well as acute physiology and chronic health evaluation (APACHE Ⅱ) were independent risk factors for predicting MODS in patients with severe sepsis.Lactate/albumin ratios between MODS group and non-MODS group, death group and non-death group were both significantly different (both P<0.05);lactate/albumin ratio was correlated with APACHE Ⅱand PaO2/FiO2, the higher the APACHE Ⅱ score and the lower the PaO2/FiO2, the higher of lactate/albumin ratio.The area under the receiver operating curve (ROC curve) analysis showed that incidence and mortality of MODS on the first day of admission predicted by lactate /albumin ratio were 0.85 and 0.84 respectively;sensitivity, specificity, positive predictive value, and negative predictive value of occurrence of MODS predicted by lactate /albumin ratio>1.735 were 80.00%, 79.17%, 82.67%, and 75.92% respectively, sensitivity, specificity, positive predictive value, and negative predictive value of mortality were 100.00%, 51.02%, 17.23%, and 100.00% respectively.Conclusion Lactate/albumin ratio level is closely correlated with incidence and mortality of MODS in patients with severe sepsis or septic shock.

Objective To construct a lentiviral vector delivering the Nicastrin (NCT) gene-targeted short hairpin RNA (shRNA) and determine gene-silencing efficiency of the vector in the human immortalized keratinocyte cell line HaCaT,and to construct a NCT gene-silenced HaCaT cell model to lay an experimental foundation for subsequently studying effects of NCT gene silencing on biological behavior of keratinocytes.Methods Three NCT gene-targeted shRNAs were designed and inserted into the pGLV3/ H1/GFP + Puro vector to construct three recombinant plasmids,which were then confirmed by sequencing.Recombinant plasmids combined with lentivirus packaging plasmids were co-transfected into 293T cells to obtain lentivirus particles,and the virus titer was determined.Cultured HaCaT cells were divided into 3 groups:blank group receiving no treatment,negative control group infected with the empty vector LV3-shNC,interference groups infected with lentivirus NCT-shRNA1,-shRNA2,-shRNA3,respectively.Flow cytometry was performed to determine transfection efficiency,and real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were conducted to determine efficiency of target gene silencing in HaCaT cells,so as to select the most efficient interference sequence.Results Sequencing analysis indicated that recombinant lentiviral vector NCT-shRNA was constructed successfully.After co-transfection of recombinant plasmids and lentivirus packaging plasmids into 293T cells,the titer of recombinant lentivirus particles was about 109 TU/ml.Flow cytometry showed that the transfection efficiency was greater than 95％.qRT-PCR revealed that the NCT mRNA expression was obviously down-regulated in the interference group compared with the negative control group,and NCT-shRNA1 was the most efficient sequence with interference efficiency being 75％.Western blot analysis showed that the inhibition rate of NCT protein expression was 71.7％ in the shRNA1 group compared with the negative control group.Conclusion The most efficient NCT-shRNA interference sequence is screened out,and the recombinant lentiviral vector NCT-shRNA and an NCT gene-silenced HaCaT cell model are both constructed successfully.

Objective:To explore the clinical value of lobar lung transplantation for end-stage lung disease patients in organ donation era.Methods:Clinical data were analyzed retrospectively for 14 cases with lobar lung transplantations between January 2016 and December 2019, including clinical outcomes and postoperative complications.Results:Eleven cases(78.6 %)had a positive etiology examination in bronchial secretion or tissue culture. There were unilateral lobar lung transplantation (n=2), bilateral lobar lung transplantation(n=2)and unilateral lobar lung transplantation and contralateral lung transplantation(n=10). Intra-operative ECMO(n=11)postoperative ECMO(n=5)were required. All survived during a 30-day perioperative period. The median time of postoperative ECMO was 1(1~11)day, the median time of extubation 4.5(0~182)day and the median time of stay in ICU 11(2~186)day. Re-operation was required for 1 case due to active bleeding in thoracic cavity. There were 10 survivors and 4 deaths. The causes of death were bronchus fistula(n=2), pulmonary infection(n=1)and renal failure(n=1)respectively.Conclusions:Lobar lung transplantation is efficacious for selected patients with end-stage lung disease.